NZ605714B - Broccoli hybrid rx 05900097 and parents thereof - Google Patents
Broccoli hybrid rx 05900097 and parents thereofInfo
- Publication number
- NZ605714B NZ605714B NZ605714A NZ60571413A NZ605714B NZ 605714 B NZ605714 B NZ 605714B NZ 605714 A NZ605714 A NZ 605714A NZ 60571413 A NZ60571413 A NZ 60571413A NZ 605714 B NZ605714 B NZ 605714B
- Authority
- NZ
- New Zealand
- Prior art keywords
- plant
- broccoli
- hybrid
- seed
- line
- Prior art date
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Abstract
Patent No. 605714 Disclosed is a broccoli plant comprising at least a first set of the chromosomes of broccoli line BRM-53-3950CMS or broccoli line BRM-53-3924SI, a sample of seed of said lines having been deposited under ATCC Accession Number PTA-12479 and ATCC Accession Number PTA-12485, respectively. vely.
Description
SEM
APPLICATION FOR LETTERS PATENT
BROCCOLI HYBRID RX 05900097 AND PARENTS THEREOF
Franciscus van den Bosch and Meinardus Boon
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims the priority of U.S. Provisional Appl. Ser. No.
61/593,243, filed January 31, 2012, and U.S. Regular Appli. Ser. No. 13/558,337, filed 25
July 2012, the entire disclosure of both of which are incorporated herein by reference.
FIELD OF THE INVENTION
The present invention relates to the field of plant breeding and, more
specifically, to the development of broccoli hybrid RX 05900097 and the parent broccoli
lines BRM3950CMS and BRM3924SI.
BACKGROUND OF THE INVENTION
The goal of vegetable breeding is to combine various desirable traits in a
single variety/hybrid. Such desirable traits may include any trait deemed beneficial by a
grower and/or consumer, including greater yield, resistance to insects or disease, tolerance to
environmental stress, and nutritional value.
Breeding techniques take advantage of a plant’s method of pollination. There
are two general methods of pollination: a plant self-pollinates if pollen from one flower is
transferred to the same or another flower of the same plant or plant variety. A plant cross-
pollinates if pollen comes to it from a flower of a different plant variety.
Plants that have been self-pollinated and selected for type over many
generations become homozygous at almost all gene loci and produce a uniform population of
true breeding progeny, a homozygous plant. A cross between two such homozygous plants
of different genotypes produces a uniform population of hybrid plants that are heterozygous
for many gene loci. Conversely, a cross of two plants each heterozygous at a number of loci
produces a population of hybrid plants that differ genetically and are not uniform. The
resulting non-uniformity makes performance unpredictable.
The development of uniform varieties requires the development of
homozygous inbred plants, the crossing of these inbred plants, and the evaluation of the
crosses. Pedigree breeding and recurrent selection are examples of breeding methods that
have been used to develop inbred plants from breeding populations. Those breeding methods
combine the genetic backgrounds from two or more plants or various other broad-based
sources into breeding pools from which new lines and hybrids derived therefrom are
developed by selfing and selection of desired phenotypes. The new lines and hybrids are
evaluated to determine which of those have commercial potential.
SUMMARY OF THE INVENTION
In one aspect, the present invention provides a broccoli plant of the hybrid
designated RX 05900097 the broccoli line BRM3950CMS or broccoli line BRM
3924SI. Also provided are broccoli plants having all the physiological and morphological
characteristics of such a plant. Parts of these broccoli plants are also provided, for example,
including pollen, an ovule, a floret, a head, and a cell of the plant.
In another aspect of the invention, a plant of broccoli hybrid RX 05900097
and/or broccoli lines BRM3950CMS and BRM3924SI comprising an added heritable
trait is provided. The heritable trait may comprise a genetic locus that is, for example, a
dominant or recessive allele. In one embodiment of the invention, a plant of broccoli hybrid
RX 05900097 and/or broccoli lines BRM3950CMS and BRM3924SI is defined as
comprising a single locus conversion. In specific embodiments of the invention, an added
genetic locus confers one or more traits such as, for example, herbicide tolerance, insect
resistance, disease resistance, and modified carbohydrate metabolism. In further
embodiments, the trait may be conferred by a naturally occurring gene introduced into the
genome of a line by backcrossing, a natural or induced mutation, or a transgene introduced
through genetic transformation techniques into the plant or a progenitor of any previous
generation thereof. When introduced through transformation, a genetic locus may comprise
one or more genes integrated at a single chromosomal location.
The invention also concerns the seed of broccoli hybrid RX 05900097 and/or
broccoli lines BRM3950CMS and BRM3924SI. The broccoli seed of the invention
may be provided, in particular embodiments, as an essentially homogeneous population of
broccoli seed of broccoli hybrid RX 05900097 and/or broccoli lines BRM3950CMS and
BRM3924SI. Essentially homogeneous populations of seed are generally free from
substantial numbers of other seed. Therefore, seed of hybrid RX 05900097 and/or broccoli
lines BRM3950CMS and BRM3924SI may be provided, in certain embodiments of
the invention, as forming at least about 97% of the total seed, including at least about 98%,
99% or more of the seed. The seed population may be separately grown to provide an
essentially homogeneous population of broccoli plants designated RX 05900097 and/or
broccoli lines BRM3950CMS and BRM3924SI.
In yet another aspect of the invention, a tissue culture of regenerable cells of a
broccoli plant of hybrid RX 05900097 and/or broccoli lines BRM3950CMS and BRM-
53-3924SI is provided. The tissue culture will preferably be capable of regenerating broccoli
plants capable of expressing all of the physiological and morphological characteristics of the
starting plant, and of regenerating plants having substantially the same genotype as the
starting plant. Examples of some of the physiological and morphological characteristics of
the hybrid RX 05900097 and/or broccoli lines BRM3950CMS and BRM3924SI
include those traits set forth in the tables herein. The regenerable cells in such tissue cultures
may be derived, for example, from embryos, meristems, cotyledons, pollen, leaves, anthers,
roots, root tips, pistils, flowers, seed and stalks. Still further, the present invention provides
broccoli plants regenerated from a tissue culture of the invention, the plants having all the
physiological and morphological characteristics of hybrid RX 05900097 and/or broccoli lines
BRM3950CMS and BRM3924SI.
In still yet another aspect of the invention, processes are provided for
producing broccoli seeds, plants and parts thereof, which processes generally comprise
crossing a first parent broccoli plant with a second parent broccoli plant, wherein at least one
of the first or second parent broccoli plants is a plant of broccoli line BRM3950CMS or
broccoli line BRM3924SI. These processes may be further exemplified as processes for
preparing hybrid broccoli seed or plants, wherein a first broccoli plant is crossed with a
second broccoli plant of a different, distinct genotype to provide a hybrid that has, as one of
its parents, a plant of broccoli line BRM3950CMS or broccoli line BRM3924SI. In
these processes, crossing will result in the production of seed. The seed production occurs
regardless of whether the seed is collected or not.
In one embodiment of the invention, the first step in “crossing” comprises
planting seeds of a first and second parent broccoli plant, often in proximity so that
pollination will occur for example, mediated by insect vectors. Alternatively, pollen can be
transferred manually. Where the plant is self-pollinated, pollination may occur without the
need for direct human intervention other than plant cultivation.
A second step may comprise cultivating or growing the seeds of first and
second parent broccoli plants into plants that bear flowers. A third step may comprise
preventing self-pollination of the plants, such as by emasculating the flowers (i.e., killing or
removing the pollen).
A fourth step for a hybrid cross may comprise cross-pollination between the
first and second parent broccoli plants. Yet another step comprises harvesting the seeds from
at least one of the parent broccoli plants. The harvested seed can be grown to produce a
broccoli plant or hybrid broccoli plant.
The present invention also provides the broccoli seeds and plants produced by
a process that comprises crossing a first parent broccoli plant with a second parent broccoli
plant, wherein at least one of the first or second parent broccoli plants is a plant of broccoli
hybrid RX 05900097 and/or broccoli lines BRM3950CMS and BRM3924SI. In one
embodiment of the invention, broccoli seed and plants produced by the process are first
generation (F ) hybrid broccoli seed and plants produced by crossing a plant in accordance
with the invention with another, distinct plant. The present invention further contemplates
plant parts of such an F hybrid broccoli plant, and methods of use thereof. Therefore, certain
exemplary embodiments of the invention provide an F hybrid broccoli plant and seed
thereof.
In still yet another aspect, the present invention provides a method of
producing a plant derived from hybrid RX 05900097 and/or broccoli lines BRM
3950CMS and BRM3924SI, the method comprising the steps of: (a) preparing a progeny
plant derived from hybrid RX 05900097 and/or broccoli lines BRM3950CMS and BRM-
53-3924SI, wherein said preparing comprises crossing a plant of the hybrid RX 05900097
and/or broccoli lines BRM3950CMS and BRM3924SI with a second plant; and (b)
crossing the progeny plant with itself or a second plant to produce a seed of a progeny plant
of a subsequent generation. In further embodiments, the method may additionally comprise:
(c) growing a progeny plant of a subsequent generation from said seed of a progeny plant of a
subsequent generation and crossing the progeny plant of a subsequent generation with itself
or a second plant; and repeating the steps for an additional 3-10 generations to produce a
plant derived from hybrid RX 05900097 and/or broccoli lines BRM3950CMS and BRM-
53-3924SI. The plant derived from hybrid RX 05900097 and/or broccoli lines BRM
3950CMS and BRM3924SI may be an inbred line, and the aforementioned repeated
crossing steps may be defined as comprising sufficient inbreeding to produce the inbred line.
In the method, it may be desirable to select particular plants resulting from step (c) for
continued crossing according to steps (b) and (c). By selecting plants having one or more
desirable traits, a plant derived from hybrid RX 05900097 and/or broccoli lines BRM
3950CMS and BRM3924SI is obtained which possesses some of the desirable traits of
the line/hybrid as well as potentially other selected traits.
In certain embodiments, the present invention provides a method of producing
food or feed comprising: (a) obtaining a plant of broccoli hybrid RX 05900097 and/or
broccoli lines BRM3950CMS and BRM3924SI, wherein the plant has been
cultivated to maturity, and (b) collecting tissue of the plant.
In still yet another aspect of the invention, the genetic complement of broccoli
hybrid RX 05900097 and/or broccoli lines BRM3950CMS and BRM3924SI is
provided. The phrase “genetic complement” is used to refer to the aggregate of nucleotide
sequences, the expression of which sequences defines the phenotype of, in the present case, a
broccoli plant, or a cell or tissue of that plant. A genetic complement thus represents the
genetic makeup of a cell, tissue or plant, and a hybrid genetic complement represents the
genetic make up of a hybrid cell, tissue or plant. The invention thus provides broccoli plant
cells that have a genetic complement in accordance with the broccoli plant cells disclosed
herein, and plants, seeds and plants containing such cells.
Plant genetic complements may be assessed by genetic marker profiles, and by
the expression of phenotypic traits that are characteristic of the expression of the genetic
complement, e.g., isozyme typing profiles. It is understood that hybrid RX 05900097 and/or
broccoli lines BRM3950CMS and BRM3924SI could be identified by any of the
many well known techniques such as, for example, Simple Sequence Length Polymorphisms
(SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990), Randomly Amplified
Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence
Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction
(AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically
incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms
(SNPs) (Wang et al., Science, 280:1077-1082, 1998).
In still yet another aspect, the present invention provides hybrid genetic
complements, as represented by broccoli plant cells, tissues, plants, and seeds, formed by the
combination of a haploid genetic complement of a broccoli plant of the invention with a
haploid genetic complement of a second broccoli plant, preferably, another, distinct broccoli
plant. In another aspect, the present invention provides a broccoli plant regenerated from a
tissue culture that comprises a hybrid genetic complement of this invention.
In still yet another aspect, the invention provides a method of determining the
genotype of a plant of broccoli hybrid RX 05900097 and/or broccoli lines BRM
3950CMS and BRM3924SI comprising detecting in the genome of the plant at least a
first polymorphism. The method may, in certain embodiments, comprise detecting a plurality
of polymorphisms in the genome of the plant. The method may further comprise storing the
results of the step of detecting the plurality of polymorphisms on a computer readable
medium. The invention further provides a computer readable medium produced by such a
method.
Any embodiment discussed herein with respect to one aspect of the invention
applies to other aspects of the invention as well, unless specifically noted.
The term “about” is used to indicate that a value includes the standard
deviation of the mean for the device or method being employed to determine the value. The
use of the term “or” in the claims is used to mean “and/or” unless explicitly indicated to refer
to alternatives only or the alternatives are mutually exclusive. When used in conjunction
with the word “comprising” or other open language in the claims, the words “a” and “an”
denote “one or more,” unless specifically noted otherwise. The terms “comprise,” “have”
and “include” are open-ended linking verbs. Any forms or tenses of one or more of these
verbs, such as “comprises,” “comprising,” “has,” “having,” “includes” and “including,” are
also open-ended. For example, any method that “comprises,” “has” or “includes” one or
more steps is not limited to possessing only those one or more steps and also covers other
unlisted steps. Similarly, any plant that “comprises,” “has” or “includes” one or more traits is
not limited to possessing only those one or more traits and covers other unlisted traits.
Other objects, features and advantages of the present invention will become
apparent from the following detailed description. It should be understood, however, that the
detailed description and any specific examples provided, while indicating specific
embodiments of the invention, are given by way of illustration only, since various changes
and modifications within the spirit and scope of the invention will become apparent to those
skilled in the art from this detailed description.
DETAILED DESCRIPTION OF THE INVENTION
The invention provides methods and compositions relating to plants, seeds and
derivatives of broccoli hybrid RX 05900097 and/or broccoli lines BRM3950CMS and
BRM3924SI. The hybrid RX 05900097 was produced by the cross of parent lines BRM-
53-3950CMS and BRM3924SI. The parent lines show uniformity and stability within
the limits of environmental influence. By crossing the parent lines, uniform plants of hybrid
RX 05900097 can be obtained.
Hybrid RX 05900097 has a Raised Head and has clean stem for ease of
harvest. The variety matures relatively uniform so that less harvest cuts are required
compared to standard broccoli varieties. In uniform conditions even single (machine) harvest
is an option.
A. Origin and Breeding History of Broccoli Hybrid RX 05900097
The parents of hybrid RX 05900097 are BRM3950CMS and BRM
3924SI.
Parent line BRM3950CMS, also known as BRM 53-3950CMS, was
created by the following breeding steps:
FEMALE BRM3950CMS : MSC100 PMMPPMN-3.1.2.
F2 PLH42/MRD6/M84(520135) X F2
PLH2546/PLH33//MAOM/NJAECB(520166)
F1 PMMPPMN
(521155)
F2 PMMPPMN-3
(530177)
F3 PMMPPMN-3.1
(540123)
F4 PMMPPMN-3.1.2
(550445)
F5 PMMPPMN-3.1.2.x
(560421/560422)
SH 29823 X F8 PMMPPMN-
3.1.2.4.1.2.1
BRM3950CMS
Parent line BRM3924SI, also known as BRM 53-3924SI, was created by
the following breeding steps:
MALE BRM 53-3924 SI : F8 PLH42/MA2-5.1.1.1.1.3.2.
F6 PLH42 (500044) X Fn MA2 943550 (500266)
F1 PLH42/MA2(501034)
F2 PLH42/MA2-5(511007)
F3 PLH42/MA2-5.1(522034)
F4 PLH42/MA2-5.1.1(532179)
F5 PLH42/MA2-5.1.1.1(542173)
F6 PLH42/MA2 5.1.1.1.1(550429)
F7 PLH42/MA2-5.1.1.1.1.3(560341)
F8 PLH42/MA2-5.1.1.1.1.3.2(570375)
BRM3924SI
B. Physiological and Morphological Characteristics of Broccoli Hybrid RX
05900097, Broccoli Line BRM3950CMS and Broccoli Line BRM3924SI
In accordance with one aspect of the present invention, there is provided a
plant having the physiological and morphological characteristics of broccoli hybrid RX
05900097 and the parent lines thereof. A description of the physiological and morphological
characteristics of such plants is presented in Tables 1-3.
Table 1: Physiological and Morphological Characteristics of Hybrid RX 05900097
Characteristic RX 05900097 Sibsey
A Region of Adaptation NW Europe NW Europe
B Maturity: fall planted
days from direct seeding to 50%
no direct seeding no direct seeding
harvest
days from transplanting to 50%
59 51
harvest
19/07/2011 19/07/2011
transplant date
Maturity: fall planted
length of harvest period in days
13-Sep-11 4-Sep-11
first harvest date
16-Sep-11 8-Sep-11
last harvest date
harvest season (main crop at 50%
fall fall
harvest)
early
very early
time of harvest maturity (50% of
(Galaxy, Packman,
(Earlyman, Primor)
plants)
Scorpio)
early**
time of beginning of flowering (50%
medium**
(Clipper, Southern
of plants with at least 10% flowers)
(Coaster, Cruiser)
Comet)
**choice for UPOV TG only
C Seedling
medium green medium green
cotyledon color
RHS Color Chart value for seedling
137B 137B
cotyledon color
weak weak
cotyledon anthocyanin
intermediate strong
hypocotyl anthocyanin
D Plant
plant height in centimeters (from soil
57.6 cm 54.1 cm
line to top of leaves)
head height in centimeters (from soil
49.1 cm 57.4 cm
line to top of head)
medium medium
height (at harvest maturity)
(Coaster) (Coaster)
one one
(Ramoso Calabrese, (Ramoso Calabrese,
number of stems
Shogun) Shogun)
Characteristic RX 05900097 Sibsey
medium many
branches
intermediate compact
habit
fresh market fresh market
market class
annual annual
life cycle
first generation hybrid first generation hybrid
type of variety
E Leaves
outer leaves: number of leaves per
18
plant (at harvest)
outer leaves: width (at midpoint of
.5 cm 16.7 cm
plant including petiole)
medium narrow
leaf: width
(Buccaneer, Green Belt) (Arcadia, Brigadeer)
outer leaves: length (at midpoint of
42.1 cm 34.1 cm
plant including petiole)
17.7 cm 13.9 cm
outer leaves: petiole length
medium
short
(Emperor, Ramoso
petiole: length
(High Sierra, Padovano)
Calabrese)
2:1 2:1
outer leaves: leaf ratio - length/width
petiolate petiolate
outer leaves: leaf attachment
intermediate intermediate
outer leaves: wax presence
few medium
leaf: number of lobes
(Early White Sprouting) (Coaster, Topper)
outer leaves: foliage color (with wax,
grey-green grey-green
if present)
outer leaves: foliage color (with wax,
189A 189A
if present; RHS Color Chart value)
grey green grey green
leaf blade: color
(Bishop) (Bishop)
medium
leaf blade: intensity of color
absent absent
leaf blade: anthocyanin coloration
(Claudia, Embassy) (Claudia, Embassy)
weak
(Beaufort, Early Pack,
leaf blade: undulation of margin
Laser, Paladin)
weak weak
leaf blade: dentation of margin
(Galaxy) (Galaxy)
elliptic elliptic
outer leaves: leaf shape
blunt blunt
outer leaves: leaf base
Characteristic RX 05900097 Sibsey
blunt blunt
outer leaves: leaf apex
slightly wavy straight
outer leaves: leaf margins
thin thin
outer leaves: leaf veins
not raised not raised
outer leaves: midrib
absent or very weak absent or very weak
leaf blade: blistering
(Buccaneer, Colibri) (Buccaneer, Colibri)
semi-erect semi-erect
outer leaves: attitude (leaf angle from
(35-55 degrees) (35-55 degrees)
ground)
semi-erect semi-erect
leaf: attitude (at beginning of head
(Arcadia, Asti, Civet, (Arcadia, Asti, Civet,
formation)
Claudia) Claudia)
none weak
outer leaves: torsion of leaf tip
outer leaves: profile of upper side of
planar convex
leaf
F Head
medium medium
length of branching at base (excluding
(Capitol, Green Duke, (Capitol, Green Duke,
stem)
Perseus) Perseus)
diameter (at widest point) (at market
14.1 cm 11.8 cm
maturity)
6.9 cm 6.1 cm
depth (at market maturity)
weight, market trimmed (at market
277.4 gm 139.3 gm
maturity)
grey-green grey-green
color (at market maturity)
(Brigadeer, Galaxy) (Brigadeer, Galaxy)
medium medium
intensity of color
N189B N189B
RHS Color Chart value for head color
present
absent
(Brigadeer, Shogun,
anthocyanin coloration
(Early White Sprouting)
Viola)
very weak very weak
intensity of anthocyanin coloration
transverse broad elliptic transverse elliptic
shape (at market maturity)
(Admiral, Corvet) (Buccaneer, Futura)
domed domed
dome shape (at market maturity)
medium** small**
size (at market maturity)
(Dundee, Early Man) (Orbit, Scorpio)
**choice for US Exhibit C only
medium medium
compactness / firmness (at market
(Late Corona) (Late Corona)
maturity)
Characteristic RX 05900097 Sibsey
medium fine
surface knobbling (at market maturity)
(Southern Comet) (Apollo, Brigadeer)
fine
medium
(Auriga, Bishop, Green
texture
(Clipper, Coaster)
Top)
small small
bead size (at market maturity)
even in size even in size
flower buds (at market maturity)
anthocyanin coloration of leaf axils (at
present absent
market maturity)
anthocyanin coloration of leaf veins
absent absent
(at market maturity)
anthocyanin coloration of leaf blade
absent absent
(at market maturity)
anthocyanin coloration of entire plant
absent absent
(at market maturity)
absent absent
anthocyanin coloration of leaf petiole
(Claudia, Embassy) (Claudia, Embassy)
(at market maturity)
color of head leaves (at market
green green
maturity)
RHS Color Chart value for the color
189A 189A
of head leaves
absent absent
bracts
(Gem, Orion) (Gem, Orion)
combination; present
secondary heads (at market maturity)
(Marathon, Tribute, Late present combination
Purple Sprouting)
intermediate
prominence of secondary heads (at
weak
(Citation)
market maturity)
number of secondary heads (at market
maturity)
G Flower
yellow yellow
Color
(Brigadeer, Orion) (Brigadeer, Orion)
RHS Color Chart value for flower
4B 4A
color
green green
stalk color
RHS Color Chart value for flower
138B 138B
stalk color
present present
male sterility
(Chevalier, Montop) (Chevalier, Montop)
*These are typical values. Values may vary due to environment. Other values that are
substantially equivalent are also within the scope of the invention.
Table 2: Physiological and Morphological Characteristics of Line BRM3950CMS
Characteristic BRM3950CMS Sibsey
A Region of Adaptation NW Europe NW Europe
B Maturity: fall planted
days from direct seeding to
no direct seeding no direct seeding
50% harvest
days from transplanting to 50%
62 51
harvest
transplant date 19-Jul-11 19-Jul-11
length of harvest period in days 3 4
first harvest date 16-Sept-11 4-Sept-11
last harvest date 19-Sept-11 8-Sept-11
harvest season (main crop at
fall fall
50% harvest)
time of harvest maturity (50% medium very early
of plants) (Sumosun) (Earlyman, Primor)
time of beginning of flowering early
medium
(50% of plants with at least (Clipper, Southern
(Coaster, Cruiser)
% flowers) Comet)
C Seedling
cotyledon color medium green medium green
RHS Color Chart value for
137A 137B
seedling cotyledon color
cotyledon anthocyanin weak weak
hypocotyl anthocyanin intermediate strong
D Plant
plant height in centimeters
46.4 cm 54.1 cm
from soil line to top of leaves
head height in centimeters from
45.3 cm 57.4 cm
soil line to top of head
short medium
height at harvest maturity
(Packman, Primor) (Coaster)
Characteristic BRM3950CMS Sibsey
one one
number of stems (Ramoso Calabrese, (Ramoso Calabrese,
Shogun) Shogun)
branches few many
habit intermediate compact
market class fresh market fresh market
life cycle annual annual
type of variety inbred first generation hybrid
E Leaves
outer leaves: number of leaves
14 18
per plant (at harvest)
outer leaves: width (at midpoint
17.9 cm 16.7 cm
of plant including petiole)
narrow narrow
leaf: width
(Arcadia, Brigadeer) (Arcadia, Brigadeer)
outer leaves: length (at
midpoint of plant including 37.5 cm 34.2 cm
petiole)
short
short
leaf: length (including petiole) (Dandy Early,
(Dandy Early, Emperor)
Emperor)
outer leaves: petiole length 16.3 cm 13.9 cm
outer leaves: leaf ratio -
2:1 2:1
length/width
outer leaves: leaf attachment petiolate petiolate
outer leaves: wax presence intermediate intermediate
few medium
leaf: number of lobes
(Early White Sprouting) (Coaster, Topper)
outer leaves: foliage color (with
grey-green grey-green
wax, if present)
outer leaves: foliage color (with
wax, if present; RHS Color 189A 189A
Chart value)
Characteristic BRM3950CMS Sibsey
grey green grey green
leaf blade: color
(Bishop) (Bishop)
leaf blade: intensity of color light medium
leaf blade: anthocyanin absent absent
coloration (Claudia, Embassy) (Claudia, Embassy)
leaf blade: undulation of
absent or very weak
margin
weak weak
leaf blade: dentation of margin
(Galaxy) (Galaxy)
outer leaves: leaf shape broad elliptic elliptic
outer leaves: leaf base blunt blunt
outer leaves: leaf apex blunt blunt
outer leaves: leaf margins slightly wavy straight
outer leaves: leaf veins thin thin
outer leaves: midrib not raised not raised
absent or very weak absent or very weak
leaf blade: blistering
(Buccaneer, Colibri) (Buccaneer, Colibri)
outer leaves: attitude (leaf semi-erect semi-erect
angle from ground) (35-55 degrees) (35-55 degrees)
semi-erect
semi-erect
leaf: attitude (at beginning of
(Arcadia, Asti, Civet,
(Arcadia, Asti, Civet,
head formation)
Claudia)
Claudia)
outer leaves: torsion of leaf tip none weak
outer leaves: profile of upper
planar convex
side of leaf
F Head
medium medium
length of branching at base
(Capitol, Green Duke, (Capitol, Green Duke,
(excluding stem)
Perseus) Perseus)
diameter (at widest point)
13.1 cm 11.8 cm
(at market maturity)
depth (at market maturity) 8.6 cm 6.1 cm
Characteristic BRM3950CMS Sibsey
weight, market trimmed (at
185.0 gm 139.3 gm
market maturity)
grey-green grey-green
color (at market maturity)
(Brigadeer, Galaxy) (Brigadeer, Galaxy)
RHS Color Chart value for
N189B N189B
head color
absent
absent
anthocyanin coloration (Early White
(Early White Sprouting)
Sprouting)
transverse broad elliptic transverse elliptic
shape (at market maturity)
(Admiral, Corvet) (Buccaneer, Futura)
dome shape (at market
domed domed
maturity)
size (at market maturity) small small
(Orbit, Scorpio) (Orbit, Scorpio)
compactness / firmness (at medium medium
market maturity) (Late Corona) (Late Corona)
surface knobbling (at market coarse fine
maturity) (Perseus, Regilio) (Apollo, Brigadeer)
fine
very fine
texture (Auriga, Bishop, Green
(Viola)
Top)
bead size (at market maturity) small small
flower buds (at market
even in size even in size
maturity)
anthocyanin coloration of leaf
absent absent
axils (at market maturity)
anthocyanin coloration of leaf
absent absent
veins (at market maturity)
anthocyanin coloration of leaf
absent absent
blade (at market maturity)
anthocyanin coloration of
entire plant (at market absent absent
maturity)
Characteristic BRM3950CMS Sibsey
anthocyanin coloration of leaf absent absent
petiole (at market maturity) (Claudia, Embassy) (Claudia, Embassy)
color of head leaves (at market
green green
maturity)
color of head leaves (at market
189A 189A
maturity)
absent absent
bracts
(Gem, Orion) (Gem, Orion)
combination;
secondary heads (at market present
present combination
maturity) (Marathon, Tribute, Late
Purple Sprouting)
prominence of secondary heads intermediate intermediate
(at market maturity) (Citation) (Citation)
number of secondary heads (at
market maturity)
G Flower
yellow yellow
color
(Brigadeer, Orion) (Brigadeer, Orion)
medium dark
intensity of yellow color
(Capitol, Corvet) (Gem, Orion)
RHS Color Chart value for
4B 4A
flower color
stalk color green green
RHS Color Chart value for
138B 138B
flower stalk color
present present
male sterility
(Chevalier, Montop) (Chevalier, Montop)
*These are typical values. Values may vary due to environment. Other values that are
substantially equivalent are also within the scope of the invention.
Table 3: Physiological and Morphological Characteristics of Line BRM3924SI
Characteristic BRM3924SI Parthenon
A Region of Adaptation most regions most regions
B Maturity: fall planted
Characteristic BRM3924SI Parthenon
days from direct seeding to 50%
no direct seeding no direct seeding
harvest
days from transplanting to 50%
72 84
harvest
19-Nov-09 19-Nov-09
transplant date
length of harvest period in days
1-Feb-10 12-Feb-10
first harvest date
harvest season (main crop at 50%
fall / winter fall / winter
harvest)
early
medium
time of harvest maturity (50% of
(Galaxy, Packman,
(Sumosun)
plants)
Scorpio)
medium late
time of beginning of flowering (50%
(Coaster, Cruiser) (Shogun, Viola)
of plants with at least 10% flowers)
C Seedling
medium green medium green
cotyledon color
RHS Color Chart value for seedling
138B 138A
cotyledon color
absent absent
cotyledon anthocyanin
weak weak
hypocotyl anthocyanin
D Plant
plant height in centimeters from soil
38.77 cm 75.33 cm
line to top of head
head height in centimeters from soil
31.67 cm 29 cm
line to top of head
short tall
height at harvest maturity
(Packman, Primor) (Citation)
one one
(Ramoso Calabrese, (Ramoso Calabrese,
number of stems
Shogun) Shogun)
few few
branches
intermediate spreading
habit
fresh market fresh market
market class
annual annual
life cycle
inbred first generation hybrid
type of variety
E Leaves
outer leaves: number of leaves per
12.33 15.33
plant (at harvest)
outer leaves: width (at midpoint of
16.6 cm 27.68 cm
plant including petiole)
Characteristic BRM3924SI Parthenon
medium
narrow
(Buccaneer, Green
leaf: width
(Arcadia, Brigadeer)
Belt)
outer leaves: length (at midpoint of
27.73 cm 65.07 cm
plant including petiole)
short long
leaf: length (including petiole)
(Dandy Early, Emperor) (Green Duke, Laser)
6.53 cm 30.33 cm
outer leaves: petiole length
short
very long
petiole: length
(High Sierra, Padovano)
2:1 3:1
outer leaves: leaf ratio - length/width
petiolate petiolate
outer leaves: leaf attachment
intermediate weak
outer leaves: wax presence
medium medium
leaf: number of lobes
(Coaster, Topper) (Coaster, Topper)
outer leaves: foliage color (with wax,
grey-green grey-green
if present)
outer leaves: foliage color (with wax,
N189A N137A
if present; RHS Color Chart value)
grey green grey green
leaf blade: color
(Bishop) (Bishop)
dark medium
leaf blade: intensity of color
absent absent
leaf blade: anthocyanin coloration
(Claudia, Embassy) (Claudia, Embassy)
strong
medium
(Aikido, Marathon,
leaf blade: undulation of margin
(Citation)
Samurai)
weak weak
leaf blade: dentation of margin
(Galaxy) (Galaxy)
elliptic broad elliptic
outer leaves: leaf shape
blunt blunt
outer leaves: leaf base
blunt blunt
outer leaves: leaf apex
very wavy slightly wavy
outer leaves: leaf margins
thick thick
outer leaves: leaf veins
raised raised
outer leaves: midrib
weak absent or very weak
leaf blade: blistering
(Coaster, Gem) (Buccaneer, Colibri)
absent absent
petiole: anthocyanin coloration of leaf
(Claudia, Embassy) (Claudia, Embassy)
petiole
Characteristic BRM3924SI Parthenon
semi-erect semi-erect
outer leaves: attitude (leaf angle from
(35-55 degrees) (35-55 degrees)
ground)
horizontal
leaf: attitude (at beginning of head
(Bishop, Colonel, New horizontal
formation)
Light)
intermediate weak
outer leaves: torsion of leaf tip
outer leaves: profile of upper side of
concave planar
leaf
F Head
short medium
length of branching at base (excluding
(Brigadeer, Buccaneer, (Capitol, Green Duke,
stem)
Emperor) Perseus)
diameter (at widest point) (at market
8.98 cm 16.91 cm
maturity)
4.95 cm 7.83 cm
depth (at market maturity)
weight (market trimmed) (at market
144.47 gm 773.73 gm
maturity)
grey-green
dark green
color (at market maturity)
(Brigadeer, Galaxy)
medium dark
intensity of color
N187C 189A
RHS Color Chart value for head color
present present
(Brigadeer, Shogun, (Brigadeer, Shogun,
anthocyanin coloration
Viola) Viola)
strong very weak
intensity of anthocyanin coloration
transverse broad
transverse elliptical
elliptical (Admiral,
shape (at market maturity)
(Buccaneer, Futura)
Futura)
semi-domed semi-domed
dome shape (at market value)
large
small
(Caravel, Mercedes,
size (at market maturity)
(Orbit, Scorpio)
Packman)
short pedicels / tight / short pedicels / tight /
compactness / firmness (at market
firm firm
maturity)
(Captain) (Captain)
fine fine
surface knobbling (at market maturity)
(Apollo, Brigadeer) (Apollo, Brigadeer)
medium medium
texture
(Clipper, Coaster) (Clipper, Coaster)
medium medium
bead size (at market maturity)
Characteristic BRM3924SI Parthenon
even in size even in size
flower buds (at market maturity)
anthocyanin coloration of leaf axils (at
absent absent
market maturity)
anthocyanin coloration of leaf veins
absent absent
(at market maturity)
anthocyanin coloration of leaf blade
absent absent
(at market maturity)
anthocyanin coloration of entire plant
absent absent
(at market maturity)
anthocyanin coloration of leaf petiole
absent absent
(at market maturity)
color of head leaves (at market
green green
maturity)
RHS Color Chart value for the color
N137B N137B
of head leaves
present present
bracts
(Ramoso Calabrese) (Ramoso Calabrese)
completely absent
combination
secondary heads (at market maturity)
(Scorpio, Zeus)
intermediate
prominence of secondary heads (at
(Citation)
market maturity)
G Flower
yellow yellow
Color
(Brigadeer, Orion) (Brigadeer, Orion)
light light
intensity of yellow color
(Brigadeer) (Brigadeer)
RHS Color Chart value for flower
4C 4C
color
green green
stalk color
RHS Color Chart value for flower
144A 144A
stalk color
absent present
male sterility
(Marathon) (Chevalier, Montop)
*These are typical values. Values may vary due to environment. Other values that are
substantially equivalent are also within the scope of the invention.
C. Breeding Broccoli Plants
One aspect of the current invention concerns methods for producing seed of
broccoli hybrid RX 05900097 involving crossing broccoli lines BRM3950CMS and
BRM3924SI. Alternatively, in other embodiments of the invention, hybrid RX
05900097, line BRM3950CMS, or line BRM3924SI may be crossed with itself or
with any second plant. Such methods can be used for propagation of hybrid RX 05900097
and/or the broccoli lines BRM3950CMS and BRM3924SI, or can be used to produce
plants that are derived from hybrid RX 05900097 and/or the broccoli lines BRM
3950CMS and BRM3924SI. Plants derived from hybrid RX 05900097 and/or the
broccoli lines BRM3950CMS and BRM3924SI may be used, in certain
embodiments, for the development of new broccoli varieties.
The development of new varieties using one or more starting varieties is well
known in the art. In accordance with the invention, novel varieties may be created by
crossing hybrid RX 05900097 followed by multiple generations of breeding according to
such well known methods. New varieties may be created by crossing with any second plant.
In selecting such a second plant to cross for the purpose of developing novel lines, it may be
desired to choose those plants which either themselves exhibit one or more selected desirable
characteristics or which exhibit the desired characteristic(s) when in hybrid combination.
Once initial crosses have been made, inbreeding and selection take place to produce new
varieties. For development of a uniform line, often five or more generations of selfing and
selection are involved.
Uniform lines of new varieties may also be developed by way of double-
haploids. This technique allows the creation of true breeding lines without the need for
multiple generations of selfing and selection. In this manner true breeding lines can be
produced in as little as one generation. Haploid embryos may be produced from microspores,
pollen, anther cultures, or ovary cultures. The haploid embryos may then be doubled
autonomously, or by chemical treatments (e.g. colchicine treatment). Alternatively, haploid
embryos may be grown into haploid plants and treated to induce chromosome doubling. In
either case, fertile homozygous plants are obtained. In accordance with the invention, any of
such techniques may be used in connection with a plant of the invention and progeny thereof
to achieve a homozygous line.
Backcrossing can also be used to improve an inbred plant. Backcrossing
transfers a specific desirable trait from one inbred or non-inbred source to an inbred that lacks
that trait. This can be accomplished, for example, by first crossing a superior inbred (A)
(recurrent parent) to a donor inbred (non-recurrent parent), which carries the appropriate
locus or loci for the trait in question. The progeny of this cross are then mated back to the
superior recurrent parent (A) followed by selection in the resultant progeny for the desired
trait to be transferred from the non-recurrent parent. After five or more backcross
generations with selection for the desired trait, the progeny have the characteristic being
transferred, but are like the superior parent for most or almost all other loci. The last
backcross generation would be selfed to give pure breeding progeny for the trait being
transferred.
The plants of the present invention are particularly well suited for the
development of new lines based on the elite nature of the genetic background of the plants.
In selecting a second plant to cross with RX 05900097 and/or broccoli lines BRM
3950CMS and BRM3924SI for the purpose of developing novel broccoli lines, it will
typically be preferred to choose those plants which either themselves exhibit one or more
selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid
combination. Examples of desirable traits may include, in specific embodiments, high seed
yield, high seed germination, seedling vigor, high yield, disease tolerance or resistance, and
adaptability for soil and climate conditions. Consumer-driven traits, such as a head shape,
nutritional value, and taste are other examples of traits that may be incorporated into new
lines of broccoli plants developed by this invention.
D. Further Embodiments of the Invention
In certain aspects of the invention, plants described herein are provided
modified to include at least a first desired heritable trait. Such plants may, in one
embodiment, be developed by a plant breeding technique called backcrossing, wherein
essentially all of the morphological and physiological characteristics of a variety are
recovered in addition to a genetic locus transferred into the plant via the backcrossing
technique. The term single locus converted plant as used herein refers to those broccoli
plants which are developed by a plant breeding technique called backcrossing, wherein
essentially all of the morphological and physiological characteristics of a variety are
recovered in addition to the single locus transferred into the variety via the backcrossing
technique. By essentially all of the morphological and physiological characteristics, it is
meant that the characteristics of a plant are recovered that are otherwise present when
compared in the same environment, other than an occasional variant trait that might arise
during backcrossing or direct introduction of a transgene.
Backcrossing methods can be used with the present invention to improve or
introduce a characteristic into the present variety. The parental broccoli plant which
contributes the locus for the desired characteristic is termed the nonrecurrent or donor parent.
This terminology refers to the fact that the nonrecurrent parent is used one time in the
backcross protocol and therefore does not recur. The parental broccoli plant to which the
locus or loci from the nonrecurrent parent are transferred is known as the recurrent parent as
it is used for several rounds in the backcrossing protocol.
In a typical backcross protocol, the original variety of interest (recurrent
parent) is crossed to a second variety (nonrecurrent parent) that carries the single locus of
interest to be transferred. The resulting progeny from this cross are then crossed again to the
recurrent parent and the process is repeated until a broccoli plant is obtained wherein
essentially all of the morphological and physiological characteristics of the recurrent parent
are recovered in the converted plant, in addition to the single transferred locus from the
nonrecurrent parent.
The selection of a suitable recurrent parent is an important step for a
successful backcrossing procedure. The goal of a backcross protocol is to alter or substitute a
single trait or characteristic in the original variety. To accomplish this, a single locus of the
recurrent variety is modified or substituted with the desired locus from the nonrecurrent
parent, while retaining essentially all of the rest of the desired genetic, and therefore the
desired physiological and morphological constitution of the original variety. The choice of
the particular nonrecurrent parent will depend on the purpose of the backcross; one of the
major purposes is to add some commercially desirable trait to the plant. The exact
backcrossing protocol will depend on the characteristic or trait being altered and the genetic
distance between the recurrent and nonrecurrent parents. Although backcrossing methods are
simplified when the characteristic being transferred is a dominant allele, a recessive allele, or
an additive allele (between recessive and dominant), may also be transferred. In this instance
it may be necessary to introduce a test of the progeny to determine if the desired
characteristic has been successfully transferred.
In one embodiment, progeny broccoli plants of a backcross in which a plant
described herein is the recurrent parent comprise (i) the desired trait from the non-recurrent
parent and (ii) all of the physiological and morphological characteristics of the recurrent
parent as determined at the 5% significance level when grown in the same environmental
conditions.
New varieties can also be developed from more than two parents. The
technique, known as modified backcrossing, uses different recurrent parents during the
backcrossing. Modified backcrossing may be used to replace the original recurrent parent
with a variety having certain more desirable characteristics or multiple parents may be used
to obtain different desirable characteristics from each.
With the development of molecular markers associated with particular traits, it
is possible to add additional traits into an established germ line, such as represented here,
with the end result being substantially the same base germplasm with the addition of a new
trait or traits. Molecular breeding, as described in Moose and Mumm, 2008 (Plant
Physiology, 147: 969-977), for example, and elsewhere, provides a mechanism for
integrating single or multiple traits or QTL into an elite line. This molecular breeding-
facilitated movement of a trait or traits into an elite line may encompass incorporation of a
particular genomic fragment associated with a particular trait of interest into the elite line by
the mechanism of identification of the integrated genomic fragment with the use of flanking
or associated marker assays. In the embodiment represented here, one, two, three or four
genomic loci, for example, may be integrated into an elite line via this methodology. When
this elite line containing the additional loci is further crossed with another parental elite line
to produce hybrid offspring, it is possible to then incorporate at least eight separate additional
loci into the hybrid. These additional loci may confer, for example, such traits as a disease
resistance or a fruit quality trait. In one embodiment, each locus may confer a separate trait.
In another embodiment, loci may need to be homozygous and exist in each parent line to
confer a trait in the hybrid. In yet another embodiment, multiple loci may be combined to
confer a single robust phenotype of a desired trait.
Many single locus traits have been identified that are not regularly selected for
in the development of a new inbred but that can be improved by backcrossing techniques.
Single locus traits may or may not be transgenic; examples of these traits include, but are not
limited to, herbicide resistance, resistance to bacterial, fungal, or viral disease, insect
resistance, modified fatty acid or carbohydrate metabolism, and altered nutritional quality.
These comprise genes generally inherited through the nucleus.
Direct selection may be applied where the single locus acts as a dominant trait.
For this selection process, the progeny of the initial cross are assayed for viral resistance
and/or the presence of the corresponding gene prior to the backcrossing. Selection eliminates
any plants that do not have the desired gene and resistance trait, and only those plants that
have the trait are used in the subsequent backcross. This process is then repeated for all
additional backcross generations.
Selection of broccoli plants for breeding is not necessarily dependent on the
phenotype of a plant and instead can be based on genetic investigations. For example, one
can utilize a suitable genetic marker which is closely genetically linked to a trait of interest.
One of these markers can be used to identify the presence or absence of a trait in the offspring
of a particular cross, and can be used in selection of progeny for continued breeding. This
technique is commonly referred to as marker assisted selection. Any other type of genetic
marker or other assay which is able to identify the relative presence or absence of a trait of
interest in a plant can also be useful for breeding purposes. Procedures for marker assisted
selection are well known in the art. Such methods will be of particular utility in the case of
recessive traits and variable phenotypes, or where conventional assays may be more
expensive, time consuming or otherwise disadvantageous. Types of genetic markers which
could be used in accordance with the invention include, but are not necessarily limited to,
Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 1
8:6531-6535, 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA
Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),
Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length
Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its
entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., Science, 280:1077-
1082, 1998).
E. Plants Derived by Genetic Engineering
Many useful traits that can be introduced by backcrossing, as well as directly
into a plant, are those which are introduced by genetic transformation techniques. Genetic
transformation may therefore be used to insert a selected transgene into a plant of the
invention or may, alternatively, be used for the preparation of transgenes which can be
introduced by backcrossing. Methods for the transformation of plants that are well known to
those of skill in the art and applicable to many crop species include, but are not limited to,
electroporation, microprojectile bombardment, Agrobacterium-mediated transformation and
direct DNA uptake by protoplasts.
To effect transformation by electroporation, one may employ either friable
tissues, such as a suspension culture of cells or embryogenic callus or alternatively one may
transform immature embryos or other organized tissue directly. In this technique, one would
partially degrade the cell walls of the chosen cells by exposing them to pectin-degrading
enzymes (pectolyases) or mechanically wound tissues in a controlled manner.
An efficient method for delivering transforming DNA segments to plant cells
is microprojectile bombardment. In this method, particles are coated with nucleic acids and
delivered into cells by a propelling force. Exemplary particles include those comprised of
tungsten, platinum, and preferably, gold. For the bombardment, cells in suspension are
concentrated on filters or solid culture medium. Alternatively, immature embryos or other
target cells may be arranged on solid culture medium. The cells to be bombarded are
positioned at an appropriate distance below the macroprojectile stopping plate.
An illustrative embodiment of a method for delivering DNA into plant cells by
acceleration is the Biolistics Particle Delivery System, which can be used to propel particles
coated with DNA or cells through a screen, such as a stainless steel or Nytex screen, onto a
surface covered with target cells. The screen disperses the particles so that they are not
delivered to the recipient cells in large aggregates. Microprojectile bombardment techniques
are widely applicable, and may be used to transform virtually any plant species.
Agrobacterium-mediated transfer is another widely applicable system for
introducing gene loci into plant cells. An advantage of the technique is that DNA can be
introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact
plant from a protoplast. Modern Agrobacterium transformation vectors are capable of
replication in E. coli as well as Agrobacterium, allowing for convenient manipulations (Klee
et al., Bio-Technology, 3(7):637-642, 1985). Moreover, recent technological advances in
vectors for Agrobacterium-mediated gene transfer have improved the arrangement of genes
and restriction sites in the vectors to facilitate the construction of vectors capable of
expressing various polypeptide coding genes. The vectors described have convenient multi-
linker regions flanked by a promoter and a polyadenylation site for direct expression of
inserted polypeptide coding genes. Additionally, Agrobacterium containing both armed and
disarmed Ti genes can be used for transformation.
In those plant strains where Agrobacterium-mediated transformation is
efficient, it is the method of choice because of the facile and defined nature of the gene locus
transfer. The use of Agrobacterium-mediated plant integrating vectors to introduce DNA into
plant cells is well known in the art (Fraley et al., Bio/Technology, 3:629-635, 1985; U.S.
Patent No. 5,563,055).
Transformation of plant protoplasts also can be achieved using methods based
on calcium phosphate precipitation, polyethylene glycol treatment, electroporation, and
combinations of these treatments (see, e.g., Potrykus et al., Mol. Gen. Genet., 199:183-188,
1985; Omirulleh et al., Plant Mol. Biol., 21(3):415-428, 1993; Fromm et al., Nature,
312:791-793, 1986; Uchimiya et al., Mol. Gen. Genet., 204:204, 1986; Marcotte et al.,
Nature, 335:454, 1988). Transformation of plants and expression of foreign genetic elements
is exemplified in Choi et al. (Plant Cell Rep., 13: 344–348, 1994), and Ellul et al. (Theor.
Appl. Genet., 107:462–469, 2003).
A number of promoters have utility for plant gene expression for any gene of
interest including but not limited to selectable markers, scoreable markers, genes for pest
tolerance, disease resistance, nutritional enhancements and any other gene of agronomic
interest. Examples of constitutive promoters useful for plant gene expression include, but are
not limited to, the cauliflower mosaic virus (CaMV) P-35S promoter, which confers
constitutive, high-level expression in most plant tissues (see, e.g., Odel et al., Nature,
313:810, 1985), including in monocots (see, e.g., Dekeyser et al., Plant Cell, 2:591, 1990;
Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990); a tandemly duplicated version of
the CaMV 35S promoter, the enhanced 35S promoter (P-e35S) the nopaline synthase
promoter (An et al., Plant Physiol., 88:547, 1988), the octopine synthase promoter (Fromm et
al., Plant Cell, 1:977, 1989); and the figwort mosaic virus (P-FMV) promoter as described in
U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter (P-eFMV) where the
promoter sequence of P-FMV is duplicated in tandem, the cauliflower mosaic virus 19S
promoter, a sugarcane bacilliform virus promoter, a commelina yellow mottle virus promoter,
and other plant DNA virus promoters known to express in plant cells.
A variety of plant gene promoters that are regulated in response to
environmental, hormonal, chemical, and/or developmental signals can also be used for
expression of an operably linked gene in plant cells, including promoters regulated by (1)
heat (Callis et al., Plant Physiol., 88:965, 1988), (2) light (e.g., pea rbcS-3A promoter,
Kuhlemeier et al., Plant Cell, 1:471, 1989; maize rbcS promoter, Schaffner and Sheen, Plant
Cell, 3:997, 1991; or chlorophyll a/b-binding protein promoter, Simpson et al., EMBO J.,
4:2723, 1985), (3) hormones, such as abscisic acid (Marcotte et al., Plant Cell, 1:969, 1989),
(4) wounding (e.g., wunl, Siebertz et al., Plant Cell, 1:961, 1989); or (5) chemicals such as
methyl jasmonate, salicylic acid, or Safener. It may also be advantageous to employ organ-
specific promoters (e.g., Roshal et al., EMBO J., 6:1155, 1987; Schernthaner et al., EMBO J.,
7:1249, 1988; Bustos et al., Plant Cell, 1:839, 1989).
Exemplary nucleic acids which may be introduced to plants of this invention
include, for example, DNA sequences or genes from another species, or even genes or
sequences which originate with or are present in the same species, but are incorporated into
recipient cells by genetic engineering methods rather than classical reproduction or breeding
techniques. However, the term “exogenous” is also intended to refer to genes that are not
normally present in the cell being transformed, or perhaps simply not present in the form,
structure, etc., as found in the transforming DNA segment or gene, or genes which are
normally present and that one desires to express in a manner that differs from the natural
expression pattern, e.g., to over-express. Thus, the term "exogenous" gene or DNA is
intended to refer to any gene or DNA segment that is introduced into a recipient cell,
regardless of whether a similar gene may already be present in such a cell. The type of DNA
included in the exogenous DNA can include DNA which is already present in the plant cell,
DNA from another plant, DNA from a different organism, or a DNA generated externally,
such as a DNA sequence containing an antisense message of a gene, or a DNA sequence
encoding a synthetic or modified version of a gene.
Many hundreds if not thousands of different genes are known and could
potentially be introduced into a broccoli plant according to the invention. Non-limiting
examples of particular genes and corresponding phenotypes one may choose to introduce into
a broccoli plant include one or more genes for insect tolerance, such as a Bacillus
thuringiensis (B.t.) gene, pest tolerance such as genes for fungal disease control, herbicide
tolerance such as genes conferring glyphosate tolerance, and genes for quality improvements
such as yield, nutritional enhancements, environmental or stress tolerances, or any desirable
changes in plant physiology, growth, development, morphology or plant product(s). For
example, structural genes would include any gene that confers insect tolerance including but
not limited to a Bacillus insect control protein gene as described in WO 99/31248, herein
incorporated by reference in its entirety, U.S. Pat. No. 5,689,052, herein incorporated by
reference in its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, herein incorporated by
reference in their entirety. In another embodiment, the structural gene can confer tolerance to
the herbicide glyphosate as conferred by genes including, but not limited to Agrobacterium
strain CP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat. No.
,633,435, herein incorporated by reference in its entirety, or glyphosate oxidoreductase gene
(GOX) as described in U.S. Pat. No. 5,463,175, herein incorporated by reference in its
entirety.
Alternatively, the DNA coding sequences can affect these phenotypes by
encoding a non-translatable RNA molecule that causes the targeted inhibition of expression
of an endogenous gene, for example via antisense- or cosuppression-mediated mechanisms
(see, for example, Bird et al., Biotech. Gen. Engin. Rev., 9:207, 1991). The RNA could also
be a catalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desired endogenous
mRNA product (see for example, Gibson and Shillito, Mol. Biotech., 7:125,1997). Thus, any
gene which produces a protein or mRNA which expresses a phenotype or morphology
change of interest is useful for the practice of the present invention.
F. Definitions
In the description and tables herein, a number of terms are used. In order to
provide a clear and consistent understanding of the specification and claims, the following
definitions are provided:
Allele: Any of one or more alternative forms of a gene locus, all of which
alleles relate to one trait or characteristic. In a diploid cell or organism, the two alleles of a
given gene occupy corresponding loci on a pair of homologous chromosomes.
Backcrossing: A process in which a breeder repeatedly crosses hybrid
progeny, for example a first generation hybrid (F ), back to one of the parents of the hybrid
progeny. Backcrossing can be used to introduce one or more single locus conversions from
one genetic background into another.
Crossing: The mating of two parent plants.
Cross-pollination: Fertilization by the union of two gametes from different
plants.
Diploid: A cell or organism having two sets of chromosomes.
Emasculate: The removal of plant male sex organs or the inactivation of the
organs with a cytoplasmic or nuclear genetic factor or a chemical agent conferring male
sterility.
Enzymes: Molecules which can act as catalysts in biological reactions.
F Hybrid: The first generation progeny of the cross of two nonisogenic
plants.
Genotype: The genetic constitution of a cell or organism.
Haploid: A cell or organism having one set of the two sets of chromosomes
in a diploid.
Linkage: A phenomenon wherein alleles on the same chromosome tend to
segregate together more often than expected by chance if their transmission was independent.
Marker: A readily detectable phenotype, preferably inherited in codominant
fashion (both alleles at a locus in a diploid heterozygote are readily detectable), with no
environmental variance component, i.e., heritability of 1.
Phenotype: The detectable characteristics of a cell or organism, which
characteristics are the manifestation of gene expression.
Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer to genetic
loci that control to some degree numerically representable traits that are usually continuously
distributed.
Resistance: As used herein, the terms “resistance” and “tolerance” are used
interchangeably to describe plants that show no symptoms to a specified biotic pest,
pathogen, abiotic influence or environmental condition. These terms are also used to
describe plants showing some symptoms but that are still able to produce marketable product
with an acceptable yield. Some plants that are referred to as resistant or tolerant are only so
in the sense that they may still produce a crop, even though the plants are stunted and the
yield is reduced.
Regeneration: The development of a plant from tissue culture.
Royal Horticultural Society (RHS) color chart value: The RHS color chart
is a standardized reference which allows accurate identification of any color. A color’s
designation on the chart describes its hue, brightness and saturation. A color is precisely
named by the RHS color chart by identifying the group name, sheet number and letter, e.g.,
Yellow-Orange Group 19A or Red Group 41B.
Self-pollination: The transfer of pollen from the anther to the stigma of the
same plant.
Single Locus Converted (Conversion) Plant: Plants which are developed by
a plant breeding technique called backcrossing, wherein essentially all of the desired
morphological and physiological characteristics of a broccoli variety are recovered in
addition to the characteristics of the single locus transferred into the variety via the
backcrossing technique and/or by genetic transformation.
Substantially Equivalent: A characteristic that, when compared, does not
show a statistically significant difference (e.g., p = 0.05) from the mean.
Tissue Culture: A composition comprising isolated cells of the same or a
different type or a collection of such cells organized into parts of a plant.
Transgene: A genetic locus comprising a sequence which has been
introduced into the genome of a broccoli plant by transformation.
G. Deposit Information
A deposit of broccoli hybrid RX 05900097, and lines BRM3950CMS and
BRM3924SI, disclosed above and recited in the claims, has been made with the
American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, VA 20110-
2209. The date of the deposits made were April 3, 2012, February 2, 2012, and February 2,
2012, respectively. The accession numbers for those deposited seeds of broccoli hybrid RX
05900097, and inbred parent lines BRM3950CMS and BRM3924SI are ATCC
Accession Number PTA-12817, and ATCC Accession Number PTA-12479, and ATCC
Accession Number PTA-12485, respectively. Upon issuance of a patent, all restrictions upon
the deposits will be removed, and the deposits are intended to meet all of the requirements of
37 C.F.R. §1.801-1.809. The deposits will be maintained in the depository for a period of 30
years, or 5 years after the last request, or for the effective life of the patent, whichever is
longer, and will be replaced if necessary during that period.
Although the foregoing invention has been described in some detail by way of
illustration and example for purposes of clarity and understanding, it will be obvious that
certain changes and modifications may be practiced within the scope of the invention, as
limited only by the scope of the appended claims.
All references cited herein are hereby expressly incorporated herein by
reference.
Claims (33)
1. A broccoli plant comprising at least a first set of the chromosomes of broccoli line BRM3950CMS, a sample of seed of said line having been deposited under ATCC Accession Number PTA-12479.
2. A seed comprising at least a first set of the chromosomes of broccoli line BRM 3950CMS, a sample of seed of said line having been deposited under ATCC Accession Number PTA-12479.
3. The plant of claim 1, which is inbred.
4. The plant of claim 1, which is hybrid.
5. The seed of claim 2, which is inbred.
6. The seed of claim 2, which is hybrid.
7. The plant of claim 4, wherein the hybrid plant is broccoli hybrid RX 05900097, a sample of seed of said hybrid RX 05900097 having been deposited under ATCC Accession Number PTA-12817.
8. The seed of claim 6, defined as a seed of broccoli hybrid RX 05900097, a sample of seed of said hybrid RX 05900097 having been deposited under ATCC Accession Number PTA-12817.
9. The seed of claim 2, defined as a seed of line BRM3950CMS.
10. A plant part of the plant of claim 1.
11. The plant part of claim 10, further defined as a leaf, an ovule, a floret, pollen, a head, or a cell.
12. A tissue culture of regenerable cells of the plant of claim 1.
13. The tissue culture according to claim 12, comprising cells or protoplasts from a plant part selected from the group consisting of embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistil, flower, seed and stalks.
14. A broccoli plant regenerated from the tissue culture of claim 12.
15. A method of vegetatively propagating the plant of claim 1 comprising the steps of: (a) collecting tissue capable of being propagated from a plant according to claim (b) cultivating said tissue to obtain proliferated shoots; and (c) rooting said proliferated shoots to obtain rooted plantlets.
16. The method of claim 15, further comprising growing at least a first plant from said rooted plantlets.
17. A method of introducing a desired trait into a broccoli line comprising: (a) crossing a plant of line BRM3950CMS with a second broccoli plant that comprises a desired trait to produce F1 progeny, a sample of seed of said lines having been deposited under ATCC Accession Number PTA-12479; (b) selecting an F1 progeny that comprises the desired trait; (c) backcrossing the selected F1 progeny with a plant of line BRM3950CMS to produce backcross progeny; (d) selecting backcross progeny comprising the desired trait and the physiological and morphological characteristic of broccoli line BRM3950CMS; and (e) repeating steps (c) and (d) three or more times to produce selected fourth or higher backcross progeny that comprise the desired trait.
18. A broccoli plant produced by the method of claim 17.
19. A method of producing a plant comprising an added trait, the method comprising introducing a transgene conferring the trait into a plant of hybrid RX 05900097 or line BRM3950CMS, a sample of seed of said hybrid and line having been deposited under ATCC Accession Number PTA-12817 and ATCC Accession Number PTA- 12479, respectively.
20. A plant produced by the method of claim 19.
21. The plant of claim 1, comprising a transgene.
22. The plant of claim 21, wherein the transgene confers a trait selected from the group consisting of male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, modified fatty acid metabolism, environmental stress tolerance, modified carbohydrate metabolism and modified protein metabolism.
23. The plant of claim 1, comprising a single locus conversion.
24. The plant of claim 23, wherein the single locus conversion confers a trait selected from the group consisting of male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, modified fatty acid metabolism, environmental stress tolerance, modified carbohydrate metabolism and modified protein metabolism.
25. A method for producing a seed of a plant derived from at least one of hybrid RX 05900097 or line BRM3950CMS comprising the steps of: (a) crossing a broccoli plant of hybrid RX 05900097, or line BRM3950CMS with itself or a second broccoli plant; a sample of seed of said hybrid and lines having been deposited under ATCC Accession Number PTA-12817, and ATCC Accession Number PTA-12479, respectively; and (b) allowing seed of a hybrid RX 05900097 or line BRM3950CMS derived broccoli plant to form.
26. The method of claim 25, further comprising the steps of: (c) selfing a plant grown from said hybrid RX 05900097 or BRM3950CMS derived broccoli seed to yield additional hybrid RX 05900097 or line BRM- 53-3950CMS derived broccoli seed; (d) growing said additional hybrid RX 05900097 or line BRM3950CMS derived broccoli seed of step (c) to yield additional hybrid RX 05900097 opr line BRM3950CMS derived broccoli plants; and (e) repeating the crossing and growing steps of (c) and (d) to generate at least a first further hybrid RX 05900097 or line BRM3950CMS derived broccoli plant.
27. The method of claim 25, wherein the second broccoli plant is of an inbred broccoli line.
28. The method of claim 25, comprising crossing line BRM3950CMS with line BRM3924SI, a sample of seed of said lines having been deposited under ATCC Accession Number PTA-12479, and ATCC Accession Number PTA-12485, respectively.
29. The method of claim 26, further comprising: (f) crossing the further hybrid RX 05900097 or BRM3950CMS derived broccoli plant with a second broccoli plant to produce seed of a hybrid progeny plant.
30. A plant part of the plant of claim 7.
31. The plant part of claim 30, further defined as a leaf, a ovule, a floret, pollen, a head, or a cell.
32. A method of producing a broccoli seed comprising crossing the plant of claim 1 with itself or a second broccoli plant and allowing seed to form.
33. A method of producing a broccoli comprising: (a) obtaining a plant according to claim 1, wherein the plant has been cultivated to maturity; and (b) collecting a broccoli from the plant.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US61/593,243 | 2012-01-31 | ||
US13/558,337 | 2012-07-25 |
Publications (1)
Publication Number | Publication Date |
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NZ605714B true NZ605714B (en) | 2013-12-03 |
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